CN107688073A - A kind of detection method of phosphatidylserine content - Google Patents
A kind of detection method of phosphatidylserine content Download PDFInfo
- Publication number
- CN107688073A CN107688073A CN201711066602.7A CN201711066602A CN107688073A CN 107688073 A CN107688073 A CN 107688073A CN 201711066602 A CN201711066602 A CN 201711066602A CN 107688073 A CN107688073 A CN 107688073A
- Authority
- CN
- China
- Prior art keywords
- sample
- phosphatidylserine
- mobile phase
- detection method
- performance liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds
Abstract
The invention discloses a kind of detection method of phosphatidylserine content, it solves the problems, such as that existing detection method has that retention time is long, peak shape is bad.It passes through following steps：（I）Draw standard curve：Accurately weighing phosphatidylserine standard specimen is made into standard liquid, then carries out high performance liquid chromatography separation using condition of gradient elution；（II）Testing sample is prepared into the fluid sample for being appropriate for high performance liquid chromatography sample introduction；（III）Testing sample is separated using gradient elution high performance liquid chromatography, and described condition of gradient elution is identical with step I condition of gradient elution；（IV）Then using EISD record peak area, and the fit equation obtained according to step I obtains the phosphatidylserine concentration in sample.The present invention uses high performance liquid chromatography to coordinate evaporation photodetector to provide accurate quick, high sensitivity a detection method for the detection of the raw material or finished product.
Description
Technical field
Suction-operated is the present invention relates to the use of, such as material is separated into each component and carrys out test material by chromatography, especially
It is a kind of detection method that phosphatidylserine content is detected in raw material and finished product.
Background technology
Phosphatidylserine, due to its unique brain tonic and intelligence development effect, is gradually applied to all kinds of as new raw-food material
Among food and health products, but detection method lacks, and there is no the detections of the unified standard to its content such as country, place or industry
Method is provided that this is resulted in the material quality control containing phosphatidylserine and final product quality control aspect without unification
Foundation.
At present, the detection method of disclosed phosphatidylserine has two kinds, and the first is：2 months 2016 Shens disclosed in 3 days
Please number be：201510874442.3 Chinese invention patent application specification disclosed in " a kind of phosphatidylserine it is efficient
Liquid phase chromatography analytical method "；The first is：The middle promulgated by the State Council of Application No. 201410145181.7 disclosed in 25 days June in 2014
" a kind of detection method of phosphorus in food acyl serine content " disclosed in bright patent application specification.Both are disclosed specially
The method being related in sharp document is to carry out content detection, above-mentioned public affairs using high performance liquid chromatography combination EISD
The content for the phosphatidylserine being directed in open file in product is different, is had differences in the sample pre-treatments stage, and follow-up
Detection-phase, instrument parameter has differences.But detection method disclosed above still suffers from, and retention time is long, peak shape is bad,
The problems such as number of theoretical plate is low.It is still immature accordingly, with respect to the detection method of the content of phosphorus in food acyl serine, it is also necessary to
It is preferable to establish a kind of chromatographic peak separation, there is the high phosphatidylserine of accurate quick, high sensitivity, favorable reproducibility, the rate of recovery
Detection method.
The content of the invention
In order to overcome the shortcomings of that existing detection method has that retention time is long, peak shape is bad, the purpose of the present invention is to carry
For a kind of detection method of preferable, accurate quick, the highly sensitive acyl serine content of phosphorus chromatographic peak separation.
The technical solution adopted for the present invention to solve the technical problems is：A kind of detection side of phosphatidylserine content
Method, it is characterised in that：It passes through following steps：
（I）Draw standard curve：Phosphatidylserine standard specimen accurately is weighed, and is dissolved in chloroform/methanol solution, is configured to standard
Solution, then carries out high performance liquid chromatography separation by the standard liquid of 4-10 group gradient volumes, and described condition of gradient elution is：
Performance liquid chromatographic column is spherical silica gel normal phase column；Column temperature is 30-40 degrees Celsius；Mobile phase A：Normal hexane, isopropanol, acetic acid and
The volume ratio of triethylamine is 810:160-200:8-12:0.6-0.10；Mobile phase B：Isopropanol, water, the body of acetic acid and triethylamine
Product ratio is 860: 110-130:8-12:0.6-0.10；In 0-10 minutes, mobile phase A and B velocity ratio are 100:0；In 10-15
Minute, mobile phase A and B velocity ratio are 60:50-30；In 15-20 minutes, mobile phase A and B velocity ratio are 0:100；Then
Peak area is recorded using EISD, and peak area is made into standard curve to the concentration of each group sample, is fitted
Go out linear equation with one unknown, the chloroform volume fraction of described chloroform/methanol solution is 85-95%；
（II）Testing sample is prepared into the fluid sample for being appropriate for high performance liquid chromatography sample introduction；
（III）The ladder of testing sample, described condition of gradient elution and step I is separated using gradient elution high performance liquid chromatography
It is identical to spend elution requirement；
（IV）Then using EISD record peak area, and the fit equation that is obtained according to step I obtains
Phosphatidylserine concentration in sample.
In currently preferred aspect, in step I and III, condition of gradient elution is：Performance liquid chromatographic column is spherical
Silica normal phase column；Column temperature is 37 degrees Celsius；Mobile phase A：Normal hexane, isopropanol, the volume ratio of acetic acid and triethylamine are 810:
180:10:0.8；Mobile phase B：Isopropanol, water, the volume ratio of acetic acid and triethylamine are 860: 130:10:0.8；At 0-10 points
Clock, mobile phase A and B velocity ratio are 100:0；In 10-15 minutes, mobile phase A and B velocity ratio are 60:40；At 15-20 points
Clock, mobile phase A and B velocity ratio are 0:100；After 30 minutes, mobile phase A and B velocity ratio are 100:0.
In currently preferred aspect, in step I, accurately weigh 120mg standard specimen phosphatidylserines and be dissolved in 25mL
Volume ratio be 90/10 chloroform/methanol solvent in be made into standard liquid, ultrasound makes powder fully dissolve, and is placed in 4 DEG C of environment
Lower preservation, then with 10,15,20,25ul and 30ul standard liquid sample introductions, make standard curve, and fit unitary once
Equation.
In currently preferred aspect, in step I and III, the chromatographic column that uses be 4m × 125mm spherical silica gel just
Xiang Zhu, the flow velocity of mobile phase is 1.2mL/min.
In currently preferred aspect, in step II, described testing sample is selected from the food containing phosphatidylserine
And health products.
Currently preferred aspect, in step II, described testing sample is selected from formula milk, pressed candy and phosphatide
Acyl serine semifinished product.
In currently preferred aspect, in step II, described testing sample is prepared into suitable as follows
Close the fluid sample for carrying out high performance liquid chromatography sample introduction：Accurately repeat to weigh testing sample, added after being ground to fine powder state
Into 50ml volumetric flask, the volume ratio for adding 10ml is 90/10 chloroform/methanol solution, after vortex oscillation is uniformly dispersed, is surpassed
Sound 5min, is cooled to room temperature, complements to 50mL with chloroform/methanol solution, shakes up, through 0.45um filtering with microporous membrane, is fitted
Close the fluid sample for carrying out high performance liquid chromatography sample introduction.
In currently preferred aspect, in step iv, the phosphatidylserine concentration in described sample is according to formula：
To calculate, wherein：
X is the mass concentration of the phosphatidylserine in sample, and its unit is mg/100g；
C is the concentration of phosphatidylserine in the sample solution obtained in step IV according to standard curve, and its unit is μ g/mL；
V is the cumulative volume for the fluid sample that high performance liquid chromatography sample introduction is appropriate in step II, and its unit is mL；
W is sample volume in step II, and its unit is g；
F is the mass fraction of moisture in sample.
In currently preferred aspect, in step iv, the drift tube temperature for evaporating photodetector is 90-100 degrees Celsius,
Flow rate of carrier gas is 2-4 mL/min.
In currently preferred aspect, in step iv, evaporation photodetector selects peak within 11.5-12.0 minutes
For peak value, peak area of the peak area as phosphatidylserine is recorded.
In other aspects of the present invention, following technical scheme is additionally provided：
1st, phase composition and ratio are flowed：
According to the property of phosphatidylserine, the binary mobile phase composition and proportioning of selection are respectively mobile phase A：Normal hexane-different
Propyl alcohol-acetic acid-triethylamine（810:180:10:0.8）, Mobile phase B：Isopropanol-water-acetic acid-triethylamine（860: 130:10:
0.8）, its eluent gradient elution requirement such as table 1.From above-mentioned proportion of mobile phase and condition of gradient elution, each peak obtains preferable
Separation, i.e. the differentiation effect of different component is preferable.
The eluent gradient elution requirement of table 1
| Step | Run time, min | Mobile phase A, % | Mobile phase B, % |
| 1 | 0 | 100 | 0 |
| 2 | 10 | 60 | 40 |
| 3 | 15 | 0 | 100 |
| 4 | 20 | 100 | 0 |
At the initial stage of elution, mobile phase A is kept at high proportion being advantageous in a short time by phosphatidylserine from mixture
Separate, while avoid the separation of other compositions, reduce interference of the impurity to measurement result.In the gradient elution later stage, rapidly
Increase the ratio of Mobile phase B, other saturation lipids can be made to separate as early as possible, shorten the detection time of experiment.
2nd, chromatogram column temperature and flow rate of mobile phase：
（1）Post effect can be increased by properly increasing column temperature, and chromatogram column temperature is controlled at 37 DEG C, on the one hand lived compared with above-mentioned two in patent
30 DEG C, 35 DEG C, the disengaging time of phosphatidylserine can be shortened, on the other hand, to nutrient phosphatidylserine, 37 DEG C
Detection temperature, its nutritional ingredient will not be caused damage, with human body intake, absorb and matched with the temperature utilized, therefore, 37
Detected under the conditions of DEG C, can obtain accurate testing result.
（2）Because post effect is the function of mobile phase linear flow rate in post, different posts can be obtained using different flow velocitys and imitated, one
As for, when flow velocity reduces, the separating effect of chromatographic peak can be more preferable, but can also extend the retention time of component to be measured simultaneously,
Simultaneous peak height reduces, peak width becomes big, so as to directly result in the reduction of the detection sensitivity of phosphatidylserine.Therefore foundation
The internal diameter and experiment peak type of chromatographic column used, it is determined that the flow velocity of the present invention is 1.2ml/min.
3rd, the drift tube temperature and flow rate of carrier gas of photodetector are evaporated
Drift tube temperature is higher, and mobile phase more tends to seethe with excitement, then can gradually reduce signal to noise ratio, but temperature is too high, can make flowing
Mutually seethe with excitement, increase background noise.Therefore the selection of the temperature of drift tube of the present invention is on the basis of mobile phase is volatilized substantially, is produced
The lowest temperature of acceptable noise is low, i.e., drift tube temperature is 95 DEG C.
Flow rate of carrier gas is that in general, flow rate of carrier gas is smaller an important factor for influenceing HPLC-ELSD detection performances, is formed
Particles of solute it is bigger, scatter that the intensity of light is bigger, if flow rate of carrier gas is too low, mobile phase will volatilize not exclusively, and then increase
Background noise.Optimal flow rate of carrier gas is the lowest speed that highest response is produced on the basis of acceptable noise.Comprehensive two
Person, through experiment, it is determined that the present invention-and the detection of phosphatidylserine is directed to, the drift tube temperature of applicable evaporation photodetector is
95 DEG C, optimal flow rate of carrier gas is 3L/min.
The present invention on the basis of existing technology, is proved by test of many times, changes proportioning, the chromatographic column temperature of mobile phase
Degree and flow rate of mobile phase, so as to change the appearance time of phosphatidylserine so that retention time shortens, and obtains phosphatide
The optimal liquid phase chromatogram condition of acyl serine content measure.The drift tube temperature and carrier gas stream of evaporation photodetector are optimized simultaneously
Fast two key parameters, the changes of two parameters the retention time of component-phosphatidylserine to be measured is influenceed it is smaller, but
Signal to noise ratio is had a great influence.The present invention coordinates evaporation photodetector using high performance liquid chromatography, by optimizing high-efficient liquid phase color
Flowing phase composition, ratio and chromatogram column temperature and flow rate of mobile phase in spectral condition；And the drift tube in evaporation photodetector
The parameter such as temperature and flow rate of carrier gas, contained using external standard peak area method to determine the PS in raw material or finished product containing phosphatidylserine
Amount, accurate quick, high sensitivity a detection method is provided for the detection of raw material or finished product.
Brief description of the drawings
The present invention will be further described with reference to the accompanying drawings and examples.
Fig. 1 is the HPLC-ELSD figures of the standard sample of embodiments of the invention 1；
Fig. 2 is the HPLC-ELSD figures of the testing sample of embodiments of the invention 1；
Fig. 3 is the HPLC-ELSD figures of the testing sample of embodiments of the invention 2.
Embodiment
The present invention is further illustrated by the following examples, but not as limitation of the present invention.
Experimental condition and setting
First, reagent.Unless additionally illustrating, Beijing Chemical Plant is purchased from, such as：
1. phosphatidylserine standard items,
2. n-hexane（Chromatographically pure）,
3.2- propyl alcohol（Chromatographically pure）,
4. acetic acid（Analyze pure）,
5. triethylamine（Chromatographically pure）,
6. chloroform-methanol（90:10 analysis is pure）,
7. ultra-pure water；
2nd, instrument condition is as follows：
1. chromatographic column：LiChrosphere 100 diol（Merck）：4m×125mm（Merck）Or equivalent chromatographic column；
2. mobile phase：
Mobile phase A is normal hexane-isopropanol-acetic acid-triethylamine（810:180:10:0.8）,
Mobile phase B is isopropanol-water-acetic acid-triethylamine（860:130:10:0.8）；
3. carry out gradient elution by the condition of table 1；
4. flow velocity：1.2mL/min；
5. column temperature：37℃；
6. sample size：5μL；
7. detector parameters：ELSD (ELSD 2000, U.S.A；95 DEG C of drift tube temperature；Flow rate of carrier gas 3.0L/min).
Embodiment 1
A kind of detection method of phosphatidylserine content, it passes through following steps：
（I）Standard liquid is prepared with drawing standard curve：
Accurately weigh 120mg standard specimen phosphatidylserines（PS）It is dissolved in 25mL chloroform/methanol（90/10）Solvent in be made into
Standard liquid, ultrasound make powder fully dissolve, and are placed under 4 DEG C of environment and preserve, then with 10,15,20,25ul and 30ul standards
Solution sample introduction, make a standard curve；
（II）Sample solution is prepared：
Accurately repeat to weigh commercially available a certain amount of sample (certain producer's formula containing phosphatidylserine containing phosphatidylserine
Milk powder), it is added in 50ml volumetric flask, adds 10ml chloroform/methanol（90/10）, after vortex oscillation is uniformly dispersed, surpass
Sound 5min, is cooled to room temperature, complements to 50mL with chloroform/methanol solution, shakes up, be configured to sample liquid, is filtered through 0.45um micropores
Membrane filtration, sample introduction；
（III）Sample detection：
Standard liquid, sample solution are measured under above-mentioned specific chromatographic condition, in elution process during the reservation of sample
Between with standard sample compared to making qualitative analysis；
（IV）Done with standard liquid peak area after logarithm as ordinate, concentration of standard solution do logarithm after be abscissa, draw double
Logarithmic scale curve, sample solution concentration is checked in from double-log standard curve according to peak area, carries out quantitative analysis.By measuring
The response of sample final PS content is calculated from standard curve.Its chromatogram is shown in accompanying drawing 1, accompanying drawing 2.
In step iv, the phosphatidylserine concentration in described sample is according to formula：
To calculate, wherein：
X is the mass concentration of the phosphatidylserine in sample, and its unit is mg/100g；
C is the concentration of phosphatidylserine in the sample solution obtained in step IV according to standard curve, and its unit is μ g/mL；
V is the cumulative volume for the fluid sample that high performance liquid chromatography sample introduction is appropriate in step II, and its unit is mL；
W is sample volume in step II, and its unit is g；
Sample solution carries out high performance liquid chromatography measure, qualitative with retention time, obtains the chromatographic peak peak face of phosphatidylserine
Product, it is quantitative with reference to sample sample weighting amount.The content for calculating phosphatidylserine formula milk is 5.88g/100g.
Embodiment 2：
A kind of detection method of phosphatidylserine content, it passes through following steps：
（I）Standard liquid is prepared with drawing standard curve：With embodiment 1；
（II）Sample solution is prepared：
Accurately repeat to weigh commercially available a certain amount of sample (certain producer's phosphatidylserine pressed candy containing phosphatidylserine
Fruit), it is added to after pressed candy is first ground into fine powder state in 50ml volumetric flask, adds 10ml chloroform/methanol（90/
10）, after vortex oscillation is uniformly dispersed, ultrasonic 5min, room temperature is cooled to, complements to 50mL with chloroform/methanol solution, shake up, match somebody with somebody
Sample liquid is made, through 0.45um filtering with microporous membrane, sample introduction；
（III）Sample detection：
Standard liquid, sample solution are measured under above-mentioned specific chromatographic condition, in elution process during the reservation of sample
Between with standard sample compared to making qualitative analysis；
（IV）Done with standard liquid peak area after logarithm as ordinate, concentration of standard solution do logarithm after be abscissa, draw double
Logarithmic scale curve, sample solution concentration is checked in from double-log standard curve according to peak area, carries out quantitative analysis.By measuring
The response of sample final PS content is calculated from standard curve.Its chromatogram is shown in accompanying drawing 3.
In step iv, the phosphatidylserine concentration in described sample is according to formula：
To calculate, wherein：
X is the mass concentration of the phosphatidylserine in sample, and its unit is mg/100g；
C is the concentration of phosphatidylserine in the sample solution obtained in step IV according to standard curve, and its unit is μ g/mL；
V is the cumulative volume for the fluid sample that high performance liquid chromatography sample introduction is appropriate in step II, and its unit is mL；
W is sample volume in step II, and its unit is g；
Sample solution carries out high performance liquid chromatography measure, qualitative with retention time, obtains the chromatographic peak peak face of phosphatidylserine
Product, it is quantitative with reference to sample sample weighting amount.The content for calculating phosphatidylserine pressed candy is 9.95g/100g.
Experiment one：Precision Experiment
Two kinds of products that with the addition of phosphatidylserine that this experiment is selected in example 1 and example 2 are laboratory sample, according to this hair
Patent document conditional is detected described in bright experiment condition and text, each sample parallel determination 6 times.It determines knot
Fruit is as shown in table 2.Wherein, patent application ZL201510874442.3 uses the chromatographic condition of embodiment 1,
ZL201410145181.7 uses the chromatographic condition of embodiment.
Under 2 different condition determinations of table, Precision Experiment result
Under the conditions of never the relative standard deviation with the measured value under chromatographic condition can be seen that above three, present invention detection
The precision of method is all higher, but the precision testing result of the present invention is apparently higher than two other detection method.
Experiment two：The rate of recovery is tested
From the phosphatidyl silk ammonia raw material of certain company（PS：50%）, using soybean oil as soft capsule content auxiliary material, prepare difference and contain
The soft capsule semi-finished product of the content of amount, the content for controlling sterling PS are：5g/100g、10g/100g、15g/100g、20g/
100g and 25g/100g.Detected according to the method in the present invention, above-mentioned two patent application, every group 6 parallel, is returned
Yield is tested, as a result as shown in table 3.A milk powder product for being not added with ps raw materials is taken, adds ps raw materials, system thereto respectively
It is measured into sample of the ps contents listed by table 3.
Under 4 different condition determinations of table, Precision Experiment result
From experimental result as can be seen that the higher rate of recovery, i.e. silk containing phosphatidyl can be obtained using the chromatographic condition of the present invention
The product of propylhomoserin detects through this method, and loss is small, can obtain more accurately layer result.
Comparative example 1
The experimental method and embodiment 1 of this comparative example are essentially identical, and its difference is：Elution process and reality in step I and III
Apply example 1 to compare, difference is to be eluted using Mobile phase B all the time.As a result show in fig. 2 to be gone out within 11.532 minutes originally
Peak occurred at 7.242 minutes, and separating effect is very poor（Figure is not shown）, it is difficult to follow-up calculated by peak area.
Comparative example 2
The experimental method and embodiment 1 of this comparative example are essentially identical, and its difference is：Elution process and reality in step I and III
Apply example 1 to compare, difference is to be eluted using mobile phase A all the time.As a result show in fig. 2 to be gone out within 11.532 minutes originally
Peak occurred at 21.049 minutes, be kept completely separate needs more than 35 minutes, waste substantial amounts of eluent and time.
Claims (10)
- A kind of 1. detection method of phosphatidylserine content, it is characterised in that：It passes through following steps：（I）Draw standard curve：Phosphatidylserine standard specimen accurately is weighed, and is dissolved in chloroform/methanol solution, is configured to standard Solution, then carries out high performance liquid chromatography separation by the standard liquid of 4-10 group gradient volumes, and described condition of gradient elution is： Performance liquid chromatographic column is spherical silica gel normal phase column；Column temperature is 30-40 degrees Celsius；Mobile phase A：Normal hexane, isopropanol, acetic acid and The volume ratio of triethylamine is 810:160-200:8-12:0.6-0.10；Mobile phase B：Isopropanol, water, the body of acetic acid and triethylamine Product ratio is 860: 110-130:8-12:0.6-0.10；In 0-10 minutes, mobile phase A and B velocity ratio are 100:0；In 10-15 Minute, mobile phase A and B velocity ratio are 60:50-30；In 15-20 minutes, mobile phase A and B velocity ratio are 0:100；Then Peak area is recorded using EISD, and peak area is made into standard curve to the concentration of each group sample, is fitted Go out linear equation with one unknown, the chloroform volume fraction of described chloroform/methanol solution is 85-95%；（II）Testing sample is prepared into the fluid sample for being appropriate for high performance liquid chromatography sample introduction；（III）The ladder of testing sample, described condition of gradient elution and step I is separated using gradient elution high performance liquid chromatography It is identical to spend elution requirement；（IV）Then using EISD record peak area, and the fit equation that is obtained according to step I obtains Phosphatidylserine concentration in sample.
- A kind of 2. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step I and In III, condition of gradient elution is：Performance liquid chromatographic column is spherical silica gel normal phase column；Column temperature is 37 degrees Celsius；Mobile phase A：Just Hexane, isopropanol, the volume ratio of acetic acid and triethylamine are 810:180:10:0.8；Mobile phase B：Isopropanol, water, acetic acid and three second The volume ratio of amine is 860: 130:10:0.8；In 0-10 minutes, mobile phase A and B velocity ratio are 100:0；In 10-15 minutes, Mobile phase A and B velocity ratio are 60:40；In 15-20 minutes, mobile phase A and B velocity ratio are 0:100；After 30 minutes, Mobile phase A and B velocity ratio are 100:0.
- A kind of 3. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step I, It is accurate weigh 120mg standard specimen phosphatidylserines and be dissolved in the solvent for the chloroform/methanol that 25mL volume ratio is 90/10 be made into Standard liquid, ultrasound make powder fully dissolve, and are placed under 4 DEG C of environment and preserve, then with 10,15,20,25ul and 30ul standards Solution sample introduction, standard curve is made, and fit linear equation with one unknown.
- A kind of 4. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step I and In III, the chromatographic column used is 4m × 125mm spherical silica gel normal phase column, and the flow velocity of mobile phase is 1.2mL/min.
- A kind of 5. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step II In, described testing sample is selected from food and health products containing phosphatidylserine.
- A kind of 6. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step II In, described testing sample is selected from formula milk, pressed candy and phosphatidylserine semifinished product.
- A kind of 7. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step II In, described testing sample is prepared into the fluid sample for being appropriate for high performance liquid chromatography sample introduction as follows：It is accurate True repetition weighs testing sample, is added to after being ground to fine powder state in 50ml volumetric flask, and the volume ratio for adding 10ml is 90/10 chloroform/methanol solution, after vortex oscillation is uniformly dispersed, ultrasonic 5min, room temperature is cooled to, is mended with chloroform/methanol solution Foot is shaken up to 50mL, through 0.45um filtering with microporous membrane, obtains being appropriate for the fluid sample of high performance liquid chromatography sample introduction.
- A kind of 8. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step IV In, the phosphatidylserine concentration in described sample is according to formula：To calculate, wherein：X is the mass concentration of the phosphatidylserine in sample, and its unit is mg/100g；C is the concentration of phosphatidylserine in the sample solution obtained in step IV according to standard curve, and its unit is μ g/mL；V is the cumulative volume for the fluid sample that high performance liquid chromatography sample introduction is appropriate in step II, and its unit is mL；W is sample volume in step II, and its unit is g；F is the mass fraction of moisture in sample.
- A kind of 9. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step IV In, the drift tube temperature for evaporating photodetector is 90-100 degrees Celsius, and flow rate of carrier gas is 2-4 mL/min.
- A kind of 10. detection method of phosphatidylserine content according to claim 1, it is characterised in that：In step IV In, it is peak value that evaporation photodetector selects peak within 11.5-12.0 minutes, records the peak area as phosphatidylserine Peak area.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711066602.7A CN107688073B (en) | 2017-11-02 | 2017-11-02 | Method for detecting content of phosphatidylserine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711066602.7A CN107688073B (en) | 2017-11-02 | 2017-11-02 | Method for detecting content of phosphatidylserine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107688073A true CN107688073A (en) | 2018-02-13 |
| CN107688073B CN107688073B (en) | 2020-12-04 |
Family
ID=61154034
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711066602.7A Active CN107688073B (en) | 2017-11-02 | 2017-11-02 | Method for detecting content of phosphatidylserine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107688073B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111721848A (en) * | 2019-03-21 | 2020-09-29 | 仙乐健康科技股份有限公司 | Method for measuring content of phosphatidylserine |
| CN114088850A (en) * | 2021-11-26 | 2022-02-25 | 重庆伊诺生化制品有限公司 | Method for detecting phospholipid and cholesterol in polysaccharide |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884813A (en) * | 2014-04-11 | 2014-06-25 | 内蒙古伊利实业集团股份有限公司 | Method for detecting content of phosphatidylserine in food |
| CN104931618A (en) * | 2015-06-26 | 2015-09-23 | 卢航 | Multiple detection method for six phospholipid in complex sample |
| CN105177073A (en) * | 2015-09-08 | 2015-12-23 | 威海博宇食品有限公司 | Method of preparing phosphatidylserine |
| CN105301144A (en) * | 2015-12-02 | 2016-02-03 | 烟台燕园科玛健康产业有限公司 | High-performance liquid chromatography of phosphatidylserine |
| CN105296560A (en) * | 2015-12-07 | 2016-02-03 | 杏辉天力（杭州）药业有限公司 | Preparation method of phosphatidylserine |
-
2017
- 2017-11-02 CN CN201711066602.7A patent/CN107688073B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103884813A (en) * | 2014-04-11 | 2014-06-25 | 内蒙古伊利实业集团股份有限公司 | Method for detecting content of phosphatidylserine in food |
| CN104931618A (en) * | 2015-06-26 | 2015-09-23 | 卢航 | Multiple detection method for six phospholipid in complex sample |
| CN105177073A (en) * | 2015-09-08 | 2015-12-23 | 威海博宇食品有限公司 | Method of preparing phosphatidylserine |
| CN105301144A (en) * | 2015-12-02 | 2016-02-03 | 烟台燕园科玛健康产业有限公司 | High-performance liquid chromatography of phosphatidylserine |
| CN105296560A (en) * | 2015-12-07 | 2016-02-03 | 杏辉天力（杭州）药业有限公司 | Preparation method of phosphatidylserine |
Non-Patent Citations (3)
| Title |
|---|
| SHIRO MAWATARI ET AL: "Separation of intact plasmalogens and all other phospholipids by a single run of high-performance liquid chromatography", 《ANALYTICAL BIOCHEMISTRY》 * |
| 岳虹 等: "高效液相色谱法测定奶粉中磷脂酰丝氨酸含量", 《乳业科学与技术》 * |
| 朱瑶 等: "刺参性腺中6种磷脂含量的HPLC－ELSD分析", 《大连海洋大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111721848A (en) * | 2019-03-21 | 2020-09-29 | 仙乐健康科技股份有限公司 | Method for measuring content of phosphatidylserine |
| CN114088850A (en) * | 2021-11-26 | 2022-02-25 | 重庆伊诺生化制品有限公司 | Method for detecting phospholipid and cholesterol in polysaccharide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107688073B (en) | 2020-12-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Detection of organophosphorus pesticide residues in leaf lettuce and cucumber through molecularly imprinted solid-phase extraction coupled to gas chromatography | |
| CN110286186A (en) | A kind of method and Xin Huang Pian standard finger-print and application detecting Xin Huang Pian finger-print | |
| CN107688073A (en) | A kind of detection method of phosphatidylserine content | |
| CN106501393B (en) | A method of detection hickory chick Nucleosides | |
| CN107315058A (en) | A kind of method of total ginkgoic acid in detection ginkgo biloba succi | |
| CN110133169A (en) | A kind of method and application using frusemide in LC-MS detection human plasma | |
| CN107957467A (en) | A kind of method of lysophosphatidyl choline in separation determination pharmaceutical preparation | |
| CN109632997A (en) | The extraction and measuring method of regallity glycosides B in a kind of lily medicinal material | |
| CN109100456B (en) | Method for simultaneously determining content of 3 fat-soluble vitamins in multivitamin injection | |
| CN109142585B (en) | Method for detecting isomer of sodium pantothenate | |
| CN110609107A (en) | Method for detecting aflatoxins G2, G1, B2 and B1 in radix paeoniae alba decoction pieces by using ultra-high performance liquid chromatography-mass spectrometry | |
| CN106290683B (en) | A kind of method of simple substance sugar concentration in detection Aureomycin fermentation liquor | |
| CN116609444A (en) | Method for measuring content of sodium alginate as thickener in food | |
| CN111579684B (en) | Method for measuring content of total capsaicin in capsule wall material of capsule | |
| CN108181402A (en) | The detection method of content of zearalenone in a kind of cereal | |
| CN109387579A (en) | The method for measuring levamisol in dairy products | |
| CN109752465A (en) | A method of using the content of taurine in HPLC MS measurement milk powder | |
| CN112213417A (en) | Kit and method for detecting concentration of mycophenolic acid medicine in dried blood spots | |
| CN112858531A (en) | Method for establishing HPLC (high performance liquid chromatography) characteristic spectrum of eucommia leaves and extract thereof | |
| CN115327006B (en) | Method for detecting clopidogrel isomer | |
| CN109187798A (en) | Detect that non-, propoxyl group Chinese mugwort that non-, N- ethyl Tadalafei of ground of demethyl card rolling land and hydroxypropyl demethyl Tadalafei method | |
| CN115266983B (en) | Method for measuring antifungal drugs in antibacterial agent of antibacterial product by using gas chromatography mass spectrometry | |
| CN109001337B (en) | Method for detecting polysaccharide content of mesona blume | |
| CN110045034B (en) | Method for determining content of erucyl phosphatidylcholine by high performance liquid chromatography | |
| CN109061009A (en) | A kind of measuring method of fermentation liquid underpants health acid content |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |